Assessing Tolerance to Heavy-Metal Stress in Arabidopsis thaliana Seedlings
Heavy-metal soil contamination is one of the major abiotic stress factors that, by negatively affecting plant growth and development, severely limit agricultural productivity worldwide. Plants have evolved various tolerance and detoxification strategies in order to cope with heavy-metal toxicity while ensuring adequate supply of essential micronutrients at the whole-plant as well as cellular levels. Genetic studies in the model plant Arabidopsis thaliana have been instrumental in elucidating such mechanisms. The root assay constitutes a very powerful and simple method to assess heavy-metal stress tolerance in Arabidopsis seedlings. It allows the simultaneous determination of all the standard growth parameters affected by heavy-metal stress (primary root elongation, lateral root development, shoot biomass, and chlorophyll content) in a single experiment. Additionally, this protocol emphasizes the tips and tricks that become particularly useful when quantifying subtle alterations in tolerance to a given heavy-metal stress, when simultaneously pursuing a large number of plant lines, or when testing sensitivity to a wide range of heavy metals for a single line.
Key wordsArabidopsis thaliana Chlorophyll content Heavy-metal stress tolerance Lateral root development Primary root elongation Root assay Seedlings Shoot biomass
We thank Raquel Carvalho for critical reading of the manuscript. This work was supported by Fundação para a Ciência e a Tecnologia (Grant EXPL/AGR-PRO/1013/2013 and postdoctoral fellowship SFRH/BPD/44640/2008 awarded to Estelle Remy). Funding from the research unit GREEN-it “Bioresources for Sustainability” (UID/Multi/04551/2013) is also acknowledged.
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